Mitochondrial mechanisms and signaling in manganese exposure

锰暴露中的线粒体机制和信号传导

基本信息

批准号：
10734614
负责人：
AVANTI GOKHALE
金额：
$ 35.6万
依托单位：
EMORY UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-16 至 2028-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10734614
关键词：
Acute Adult Adverse effects Affect Air Binding Binding Proteins Biochemical Biological Biological Markers Brain Cell Death Cell Line Cell model Cells Cerebrum Child Chronic Cognition Cognitive Cytoplasm Cytoplasmic Granules Diagnosis Disease Double-Stranded RNA Down-Regulation Environmental Risk Factor Exclusion Exposure to Foundations Functional disorder Genetic Genetic Predisposition to Disease Genetic Transcription Goals Homeostasis Human Impairment Individual Inductively Coupled Plasma Mass Spectrometry Industry Inflammatory Inflammatory Response Intervention Knock-out Knockout Mice Manganese Manganism Mass Spectrum Analysis Measures Metabolism Metal exposure Metals Methods Mitochondria Mitochondrial Proteins Mitochondrial RNA Modeling Molecular Molecular Target Movement Movement Disorders Mus Mutation Nerve Tissue Neurologic Neurons Occupational Organoids Oxidative Stress Parkinsonian Disorders Particulate Matter Pathogenicity Pathway interactions Pharmaceutical Preparations Phenocopy Population Predisposition Production Proteins Public Health RNA RNA Processing Reactive Oxygen Species Reporter Research Design Respiration Respiratory Chain Ribosomes Risk Role Signal Transduction Testing Time Toxic Environmental Substances Toxic effect Transcript Water brain cell brain tissue cell injury crosslink cytokine experience interest mitochondrial dysfunction mitochondrial membrane neuron loss neuroprotection neurotoxic neurotoxicity novel novel strategies overexpression pollutant protein expression proteostasis response tool

项目摘要

Summary: This application focuses on a novel pathogenic mechanism that includes metal homeostasis, mitochondrial proteostasis and onset of neurological adverse effects for cognition and movement. We have identified the mitochondrial RNA granule, as a key initiator of neuronal responses to environmental and/or genetic insults that predispose an individual to disease. This RNA processing pathway in mitochondria resides upstream of the effects that have been attributed to manganese toxicity, including reduced respiratory chain activity, mitochondrial membrane depolarization, mitochondrial oxidative stress, leading to cell death. Our overall hypothesis is that manganese accumulation in mitochondria disrupts the mitochondrial RNA granule function and induces dsRNA accumulation as part of the toxicity mechanism. This disruption leads to the accumulation of dsRNA and reduced OXPHOS function and increased oxidative stress. Thus, the mitochondrial RNA granule offers us a molecular reporter of exposure and sensitivity to manganese and a potential target for neuroprotective interventions. Specific Aims: Aim 1: Identify molecular targets of manganese in mitochondria. Aim 2: Evaluate whether manganese-induced mitochondrial dysfunction induces dsRNA accumulation and pro-inflammatory responses. Aim 3: To test to what degree impaired mitochondrial RNA granule function modulates manganese sensitivity in human brain organoids and brain from wild type and SLC30a10 KO mouse. Study Design: We will employ cell models with genetic deficiencies in the manganese efflux transporter SLC30A10, which is sufficient to induce a parkinsonian syndrome in humans, as well as acutely and chronically manganese treated cells to identify Manganese binding proteins in mitochondria and to analyze the composition of the mitochondrial RNA granule by mass spectrometry approaches. RNA granule function will be studied by analyzing the processing of the mitochondrial polycistronic mitochondrial RNA in manganese challenged cells employing molecular counting with probes directed at unprocessed junctions in the transcripts. By knockout and over-expression of RNA granule components, we will assess whether mitochondrial RNA granule dysfunction is deleterious or adaptive. We will interrogate human brain organoids and mouse brain tissue for the molecular mechanisms identified in cell line models. Given the significant potential impact of RNA granule on mitochondrial function, we will evaluate whether drugs affecting mitochondrial RNA processing and downstream metabolism are neuroprotective to excess manganese exposure or exacerbate genetic vulnerability.

概括：该应用集中于一种新型的致病机制，包括金属稳态，线粒体蛋白抑制作用和对认知和运动的神经系统不良反应的发作。我们已经确定了线粒体RNA颗粒，是对环境的神经元反应的关键引发剂和/或遗传侮辱，使人患有疾病。此RNA处理途径线粒体驻留在锰毒性的上游，包括呼吸链活性减少，线粒体膜去极化，线粒体氧化压力，导致细胞死亡。我们的总体假设是锰在线粒体中的积累干扰线粒体RNA颗粒功能并诱导DSRNA积累作为毒性的一部分机制。这种破坏导致dsRNA的积累和降低的Oxphos功能，以及氧化应激增加。因此，线粒体RNA颗粒为我们提供了一个分子记者对锰的暴露和敏感性以及神经保护干预措施的潜在靶标。具体目的：目标1：确定线粒体中锰的分子靶标。 AIM 2：评估锰诱导的线粒体功能障碍是否诱导DSRNA积累和促炎反应。目标3：测试线粒体RNA颗粒功能在多大程度上受损的锰敏感性在野生型和SLC30A10 KO小鼠的人类脑官和大脑中。研究设计：我们将在锰外排中使用具有遗传缺陷的细胞模型运输者SLC30A10，足以诱导人类帕金森氏综合症，以及急性和慢性锰处理的细胞，以鉴定线粒体中的锰结合蛋白并通过质谱方法分析线粒体RNA颗粒的组成。 RNA颗粒功能将通过分析线粒体多余体的加工来研究锰中的线粒体RNA挑战细胞，该细胞采用了分子计数，该细胞用探针定向在成绩单中未经处理的连接处。通过敲除和RNA颗粒的过表达组件，我们将评估线粒体RNA颗粒功能障碍是有害还是适应性。我们将询问人脑器官和小鼠脑组织的分子机制在细胞系模型中确定。鉴于RNA颗粒对线粒体的显着潜在影响功能，我们将评估是否影响线粒体RNA处理和下游药物代谢对过量锰暴露或加剧遗传脆弱性具有神经保护作用。